The rescue of oral development of defective-micronucleate conjugants of Paramecium tetraurelia by normal gametic nuclei

1988 ◽  
Vol 90 (2) ◽  
pp. 287-293
Author(s):  
M. F. CHAU ◽  
STEPHEN F. NG
Keyword(s):  
Sexual Reproduction ◽  
Cell Lines ◽  
Early Stage ◽  
Sexual Cycle ◽  
Paramecium Tetraurelia ◽  
Laser Microbeam ◽  
Nuclear Reorganization ◽  
Oral Development ◽  
Derivatives Of

The present study further analyses the importance of postmeiotic divisional derivatives of the micronucleus in the development of the oral apparatus of Paramecium during sexual reproduction. Cell lines possessing defective micronuclei generated by laser microbeam irradiation of the micronucleus were employed. They exhibited anomalies in nuclear reorganization and stomatogenesis in the sexual cycle. During autogamy, in some cells the micronuclear cycle terminated shortly after meiosis, resulting in the loss of all postmeiotic micronuclear derivatives. Stomatogenesis became arrested at an early stage of assembly of the oral membranelles, but the old oral apparatus was resorbed as usual, leading to the production of astomatous cells at the end of the sexual cycle. Conjugation of these cell lines with normal micronucleates rescued both nucleogenesis and stomatogenesis in the defective micronucleate conjugant, primarily as a result of transfer of the male gametic nucleus from the normal conjugant to the defective-micronucleate mate. These observations demonstrate the stomatogenic significance, in particular in the initiation of oral membranelle assembly, of the gametic nuclei during sexual reproduction. The present study also suggests the possibility of micronuclear activities in the early part of the sexual cycle affecting postzygotic nucleogenesis.

The somatic function of the micronucleus in sexual reproduction of Paramecium tetraurelia: initiation of oral membranelle assembly

1988 ◽  
Vol 90 (1) ◽  
pp. 157-166
Author(s):  
M. F. CHAU ◽  
STEPHEN F. NG
Keyword(s):  
Sexual Reproduction ◽  
Laser Irradiation ◽  
Cell Lines ◽  
Paramecium Tetraurelia ◽  
Laser Microbeam ◽  
Microbeam Irradiation ◽  
Nuclear Reorganization

In a previous study, cell lines possessing defective micronuclei, generated by laser-microbeam irradiation, gave rise to cells lacking both oral apparatus and micronuclear derivatives after autogamy. It was concluded that astomy arose as a result of degeneration of all of the meiotic products of the micronuclei after meiotic telophase II, instead of leaving one product for subsequent nuclear reorganization. The present study consolidates this conclusion by employing 15 micronucleus-defective cell lines; these were generated by laser-irradiation of the micronucleus, treatment of the cells with cis-dichlorodiamineplatinum (II), and conjugation between diploids and amicronucleates to produce haploids. A good correlation between the presence of pregametic, gametic and zygotic nuclei and the initiation of oral membranelle assembly in stomatogenesis was demonstrated in 17 cases of autogamy. Therefore, postmeiotic micronuclear activities up to the zygotic nucleus stage, in particular in the gametic stage, are crucial for the initiation of oral membranelle assembly, while premeiotic micronuclear activities are insufficient. Micronuclear genic factors are also likely to be involved in the determination of the fate of the meiotic products.

scholarly journals The role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus

1986 ◽  
Vol 86 (1) ◽  
pp. 287-303
Author(s):  
L.W. Tam ◽  
S.F. Ng
Keyword(s):  
Cell Lines ◽  
Critical Period ◽  
Buccal Cavity ◽  
Early Stage ◽  
Multiple Roles ◽  
Paramecium Tetraurelia ◽  
Laser Microbeam ◽  
Microbeam Irradiation ◽  
Recessive Mutations

Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy, in agreement with previous observations on amicronucleate cells obtained by micropipetting. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis, deciding early steps of oral membranelle assembly and affecting their subsequent patterning. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei.

5-Azacytidine affects the programming of expression of the somatic nucleus of Paramecium

Development ◽  
1989 ◽  
Vol 105 (3) ◽  
pp. 559-568 ◽  
Author(s):  
F.W. Kwok ◽  
S.F. Ng
Keyword(s):  
Sexual Reproduction ◽  
Cell Lines ◽  
Compensatory Mechanism ◽  
Paramecium Tetraurelia ◽  
Growth Depression ◽  
Somatic Nucleus ◽  
Asexual Propagation ◽  
Oral Development ◽  
Asexual Cycle

This report introduces a new system in the study of programming of genomic function during development of the somatic nucleus of Paramecium tetraurelia. Previous works have established a definite, but replaceable, role of the germ nuclei (micronuclei) in oral development in the asexual cycle; their removal from the cell generates viable amicronucleate cell lines, which characteristically suffer a transient period of growth depression marked by abnormal oral development. Such cell lines gradually recover, showing that a compensatory mechanism is activated in the absence of the germ nuclei to bring the cell back to near-normal. To test the notion that the somatic nucleus (macronucleus) is involved in this compensation, cells possessing micronuclei were treated with 5-azacytidine during sexual reproduction when new somatic nuclei develop. These cells were then propagated asexually for a number of fissions in the absence of the drug, and thereafter micronuclei were removed from them. The amicronucleate cell lines generated in this manner clearly did not suffer a depression as severe as the untreated controls did in terms of growth rate and oral development, and they recovered much sooner. This supports the notion that the somatic nucleus is the physical basis of the compensatory mechanism. This study suggests that the stomatogenic sequences in question normally become repressed in the somatic nucleus developing in sexual reproduction, and that 5-azacytidine administered to the cells at this time could alter this programme which then persists during subsequent asexual propagation. The possibility that the somatic nucleus is programmed by methylation of cytosine at the 5′ position is discussed.

Interspecific micronuclear transplantation in Paramecium: nucleogenesis and stomatogenesis in asexual and sexual reproduction

Development ◽  
1988 ◽  
Vol 103 (1) ◽  
pp. 179-191
Author(s):  
M.F. Chau ◽  
S.F. Ng
Keyword(s):  
Cell Division ◽  
Sexual Reproduction ◽  
Cell Lines ◽  
Essential Role ◽  
The Other ◽  
Sexual Cycle ◽  
Interspecific Incompatibility ◽  
Multicellular Development ◽  
Oral Development ◽  
Species Specific

The micronucleus of Paramecium plays an essential role in the development of the oral apparatus in both asexual and sexual cycles. The present study analyses this somatic function of the micronucleus by interspecific transplantation of the micronucleus between two species, P. jenningsi and P. tetraurelia. The two species are similar in nucleogenesis in the sexual cycle, in the dependence of stomatogenesis on the micronucleus and in the pattern of the oral ciliature. P. jenningsi, however, has a longer oral apparatus. Renucleated cell lines were derived from heterospecific transplantation (P. jenningsi amicronucleates implanted with micronuclei of P. tetraurelia), and also from homospecific transplantation (P. jenningsi). Both homo- and heterospecific transplants exhibited abnormal micronuclear propagation during cell division. In the sexual cycle, the heterospecific transplants exhibited more severe micronuclear anomalies, suggesting interspecific incompatibility. On the other hand, the stomatogenic consequences of the two types of transplants in the asexual and sexual cycles were similar. It is concluded that micronuclear functions, in the assembly and normal patterning of the oral ciliature in the sexual cycle, are not species-specific. However, the oral apparatuses developed by the homo- and heterospecific transplants were similar in length, and approaching that of normal P. jenningsi. Hence, even though the micronucleus is necessary for developing normal oral length, the oral length characteristic of a species is determined by species-specific nonmicronuclear factors. The present findings resemble heterospecific dermal-epidermal inductive interactions in multicellular development, with the micronucleus exerting a nonspecies-specific ‘intracellular inductive stimulus’ on the oral anarchic field to promote oral development.

Cortical positioning and the fate of oral structures during the sexual process in amicronucleate homopolar tandems of Paramecium

Development ◽  
1988 ◽  
Vol 102 (3) ◽  
pp. 587-594
Author(s):  
S.F. Ng
Keyword(s):  
Sexual Reproduction ◽  
Cell Lines ◽  
Low Frequency ◽  
Positional Information ◽  
Binary Fission ◽  
Transient Growth ◽  
Paramecium Tetraurelia ◽  
Growth Depression ◽  
Sexual Process

Paramecium tetraurelia normally resorbs the pre-existing oral apparatus (and develops a new one) during sexual reproduction. Violation of this rule was found in amicronucleate cell lines. These cell lines generated chains of two cells (homopolar tandems) at a low frequency, as a result of incomplete binary fission during a transient growth depression period following emicronucleation. In autogamous chains, the proter resorbed the pre-existing oral structures, while some of the ospisthes retained them. The oral structures in the opisthes of the chains were unusually close to the opisthes' anterior end. The ectopic location of these oral structures might account for their retention, formally understood in terms of the theory of positional information. It is suggested that nongenic factors, likely involving components of the rigid cortical matrix, are involved in the fixation of positional values.

Synthesis and Cytotoxicity Assessment of Novel 7-O- and 14-O-Derivatives of Glaucocalyxin A

2020 ◽  
Vol 20 (10) ◽  
pp. 1241-1249
Author(s):  
Hong-Chuan Liu ◽  
Li-Ming Qiao ◽  
Wei Zheng ◽  
Zhao-Bao Xiang ◽  
Hai-Sheng Chen ◽  
...  
Keyword(s):  
Acute Toxicity ◽  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Folk Medicine ◽  
A549 Cells ◽  
Cancer Cell Lines ◽  
Human Cancer Cell Lines ◽  
Derivatives Of

Background: Rabdosia japonica has been historically used in China as a popular folk medicine for the treatment of cancer, hepatitis, and gastricism. Glaucocalyxin A (GLA), an ent-kaurene diterpene isolated from Rabdosia japonica, is one of the main active ingredients showing potent inhibitory effects against several types of tumor cells. To the best of our knowledge, studies regarding the structural modification and Structure- Activity Relations (SAR) of this compound have not yet been reported. Objective: The aim of this study was to discover more potent derivatives of GLA and investigate their SAR and cytotoxicity mechanisms. Methods: Novel 7-O- and 14-O-derivatives of GLA were synthesized by condensation of acids or acyl chloride. The anti-tumor activities of these derivatives against various human cancer cell lines were evaluated in vitro by MTT assays. Apoptosis assays of compound 17 (7,14-diacylation product) were performed on A549 and HL-60 cells by flow cytometry and TUNNEL. The acute toxicity of this compound was tested on mice, at the dose of 300mg per kg body weight. Results: Seventeen novel 7-O- and 14-O-derivatives of GLA (1-17) were synthesized. These compounds showed potent cytotoxicity against the tested cancer cell lines, and almost all of them were found to be more cytotoxic than GLA and oridonin. Of the synthesized derivatives, compound 17 presented the greatest cytotoxicity, with IC50 values of 0.26μM and 1.10μM in HL-60 and CCRF-CEM cells, respectively. Furthermore, this compound induced weak apoptosis of A549 cells but showed great potential in stimulating the apoptosis of HL- 60 cells. Acute toxicity assays indicated that compound 17 is relatively safer. Conclusion: The results reported herein indicate that the synthesized GLA derivatives exhibited greater cytotoxicity against leukemia cells than against other types of tumors. In particular, 7,14-diacylation product of GLA was found to be an effective anti-tumor agent. However, the cytotoxicity mechanism of this product in A549 cells is expected to be different than that in other tumor cell lines. Further research is needed to confirm this hypothesis.

scholarly journals Probing the Intracellular Bio-Nano Interface in Different Cell Lines with Gold Nanostars

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1183
Author(s):  
Cecilia Spedalieri ◽  
Gergo Péter Szekeres ◽  
Stephan Werner ◽  
Peter Guttmann ◽  
Janina Kneipp
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Early Stage ◽  
Protein Corona ◽  
Fibroblast Cell ◽  
Ultrastructural Level ◽  
Epithelial Cell Line ◽  
Animal Cells ◽  
Gold Nanostars

Gold nanostars are a versatile plasmonic nanomaterial with many applications in bioanalysis. Their interactions with animal cells of three different cell lines are studied here at the molecular and ultrastructural level at an early stage of endolysosomal processing. Using the gold nanostars themselves as substrate for surface-enhanced Raman scattering, their protein corona and the molecules in the endolysosomal environment were characterized. Localization, morphology, and size of the nanostar aggregates in the endolysosomal compartment of the cells were probed by cryo soft-X-ray nanotomography. The processing of the nanostars by macrophages of cell line J774 differed greatly from that in the fibroblast cell line 3T3 and in the epithelial cell line HCT-116, and the structure and composition of the biomolecular corona was found to resemble that of spherical gold nanoparticles in the same cells. Data obtained with gold nanostars of varied morphology indicate that the biomolecular interactions at the surface in vivo are influenced by the spike length, with increased interaction with hydrophobic groups of proteins and lipids for longer spike lengths, and independent of the cell line. The results will support optimized nanostar synthesis and delivery for sensing, imaging, and theranostics.

scholarly journals Biguanide-Based Synthesis of 1,3,5-Triazine Derivatives with Anticancer Activity and 1,3,5-Triazine Incorporated Calcium Citrate Nanoparticles

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1028
Author(s):  
Monnaya Chalermnon ◽  
Sarocha Cherdchom ◽  
Amornpun Sereemaspun ◽  
Rojrit Rojanathanes ◽  
Tanatorn Khotavivattana
Keyword(s):  
Anticancer Activity ◽  
Cell Lines ◽  
Anticancer Agent ◽  
Drug Loading ◽  
Calcium Citrate ◽  
Colorectal Cancer Cell Lines ◽  
Triazine Derivatives ◽  
Further Development ◽  
Derivatives Of ◽  
Potential Use

Twelve derivatives of biguanide-derived 1,3,5-triazines, a promising class of anticancer agent, were synthesised and evaluated for their anticancer activity against two colorectal cancer cell lines—HCT116 and SW620. 2c and 3c which are the derivatives containing o-hydroxyphenyl substituents exhibited the highest activity with IC50 against both cell lines in the range of 20–27 µM, which is comparable to the IC50 of cisplatin reference. Moreover, the potential use of the calcium citrate nanoparticles (CaCit NPs) as a platform for drug delivery system was studied on a selected 1,3,5-triazine derivative 2a. Condition optimisation revealed that the source of citrate ions and reaction time significantly influence the morphology, size and %drug loading of the particles. With the optimised conditions, “CaCit-2a NPs” were successfully synthesised with the size of 148 ± 23 nm and %drug loading of up to 16.3%. Furthermore, it was found that the release of 2a from the synthesised CaCit-2a NPs is pH-responsive, and 2a could be control released under the acidic cancer environment. The knowledge from this study is perceptive for further development of the 1,3,5-triazine-based anticancer drugs and provide the platform for the incorporation of other drugs in the CaCit NPs in the future.

scholarly journals Cytotoxic Profiling of Annotated and Diverse Chemical Libraries Using Quantitative High-Throughput Screening

2019 ◽  
Vol 25 (1) ◽  
pp. 9-20 ◽  
Author(s):  
Olivia W. Lee ◽  
Shelley Austin ◽  
Madison Gamma ◽  
Dorian M. Cheff ◽  
Tobie D. Lee ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Large Scale ◽  
Early Stage ◽  
Cancer Cell Line ◽  
Hek 293 ◽  
Cytotoxic Compounds

Cell-based phenotypic screening is a commonly used approach to discover biological pathways, novel drug targets, chemical probes, and high-quality hit-to-lead molecules. Many hits identified from high-throughput screening campaigns are ruled out through a series of follow-up potency, selectivity/specificity, and cytotoxicity assays. Prioritization of molecules with little or no cytotoxicity for downstream evaluation can influence the future direction of projects, so cytotoxicity profiling of screening libraries at an early stage is essential for increasing the likelihood of candidate success. In this study, we assessed the cell-based cytotoxicity of nearly 10,000 compounds in the National Institutes of Health, National Center for Advancing Translational Sciences annotated libraries and more than 100,000 compounds in a diversity library against four normal cell lines (HEK 293, NIH 3T3, CRL-7250, and HaCat) and one cancer cell line (KB 3-1, a HeLa subline). This large-scale library profiling was analyzed for overall screening outcomes, hit rates, pan-activity, and selectivity. For the annotated library, we also examined the primary targets and mechanistic pathways regularly associated with cell death. To our knowledge, this is the first study to use high-throughput screening to profile a large screening collection (>100,000 compounds) for cytotoxicity in both normal and cancer cell lines. The results generated here constitute a valuable resource for the scientific community and provide insight into the extent of cytotoxic compounds in screening libraries, allowing for the identification and avoidance of compounds with cytotoxicity during high-throughput screening campaigns.

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